Cationic emulsion of cyclosporine for the management of ocular surface inflammation: a preclinical evaluation
Philippe Daull1, Laurence Feraille2, Mourad Amrane1, Pierre-Paul Elena2, Jean-Sébastien Garrigue1
1 Santen SAS, Novagali Innovation center, Evry, France; 2 Iris Pharma, La gaude, France
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A rat corneal scrapping assay and a mouse model of DED2
were used to assess the efficacy of the CKC-CE of CsA (Ikervis)
and its vehicle. Following corneal debridement (in the rat)
and twice daily dosing for five days, in vivo confocal
microscopy (IVCM) was used to both characterize and
quantify the cornea wound healing process and inflammatory
cells infiltration in the corneal stroma, respectively.
In a controlled environment DED mouse model (8-12-week-
old female C57BL6 mice), following DED induction and three
times daily dosing for 7 days (from D3 to D10), corneal
recovery was monitored by corneal fluorescein staining (CFS).
At the end of the treatment period (D10), flat mounted
corneas were used to quantify the infiltrated CD11b+ and
CD4+ positive cells to assess the anti-inflammatory properties
of the CKC-CE of CsA (Ikervis).
Dry eye disease (DED) is a complex multifactorial pathology
characterized by corneal epithelium lesions and
inflammation. The severity of these assaults is often
correlated to the severity of the disease. DED is estimated to
affect approximately 25-30 million people in the United
States. Patients with severe DED lack a convenient and
effective therapy to treat their signs (corneal fluorescein
staining (CFS) assessing keratitis), and alleviate their
symptoms and protect their ocular surface.
The aim of the present studies was to evaluate the efficacy of
an unpreserved cetalkonium chloride (CKC)-cationic emulsion
(CE) of cyclosporine (CsA; Ikervis)1 on the management of
corneal inflammation in animal models mimicking DED-
related ocular surface inflammation.
Parameters Cationic oil-in-water emulsion of CsA
White opaque to slightly translucent
Mean droplet size (
nm)a 170 (100%)
Zeta potential (mV)
b Positive (+ 40 mV)
Table 1. Summary of the physico-chemical parameters of the
cationic oil-in-water emulsion of CsA.
Note: a Mean droplet size was determined by dynamic light scattering (HPPS,
Malvern Instruments), and b zeta potential by electrophoretic mobility
measurement (Zetasizer 2000, Malvern Instruments).
In the rat scraping assay, corneal recovery was complete by
day 5. Interestingly, the number of inflammatory cells/area
within the corneal stroma was reduced following treatments
with the CKC-CE of CsA (Ikervis, 8 cell/area) vs. saline (25
In the mouse DED model, after 7 days of treatment, the CFS
score was reduced by 59% with the CKC-CE of CsA (Ikervis,
CFS score at D3 before treatment: 12.1 ± 1.7; vs D10: 5.5 ±
2.0). The beneficial effect of the CKC-CE vehicle (Ikervis
vehicle) itself on keratitis was also clearly evidenced by its
better performance over the 1% methylprednisolone eye
drop, -36%, vs. -28%, respectively.
A slight reduction in CD11b+ and CD4+ cells was also observed
for the CKC-CE of CsA (Ikervis) in cornea when compared to
the untreated group. An enumeration of CD4+ cells into the
conjunctiva is currently in progress.
These studies indicate that Ikervis, a cationic emulsion of
cyclosporine (0.1%) is an effective formulation for the
management of corneal epithelium and stroma inflammation.
In addition, it performed better than a potent gluco-
corticosteroid (1% methylprednisolone)4. Hence, Ikervis, a
CKC-CE of CsA, may represent a promising new treatment
strategy for the management corneal inflammation in DED
Note: In March 2015 Ikervis® (IKERVIS 1 mg/mL eye drops,
emulsion) Marketing Authorization was granted by the
European Commission based on EMA’s January 2015 positive
Ikervis®, once a day at bed time, is indicated for "Treatment of
severe keratitis in adult patients with dry eye disease, which
has not improved despite treatment with tear substitutes".
Figure 1. CFS staining was used to characterize the scraped
region; at day 1, immediately after corneal scraping (A), and at
day 5 to evaluate corneal recovery (B).
Figure 3. Corneal fluorescein staining over time of mice
(n=10/group) placed in a controlled environmental chamber
and subjected to treatments (from D3 to D10). Baseline:
before DED induction; D3, post DED induction & before
treatment. Data are mean ± sd.
Figure 4. (A) CD11b+, and (B) CD4+ cells count in flat mounted
corneas (n=10) of mice treated with Ikervis at D10.
Figure 2. Following corneal scraping and five days treatment
with the different eye drops the cornea was observed by in
vivo confocal microscopy (IVCM) and compared to baseline
values from unscraped saline-treated rats. (A) IVCM scores at
day 5 were determined according to Baudouin et al. scale3. (B)
Inflammatory cells count in the corneal stroma at day 5. Rats
per group, n=6. (mean ± sem).